Blood processing apparatus and method for detoxifying bacterial lipopolysaccharide in vivo

Abstract

A detoxification method for treating sepsis, microbial infections, and other inflammatory conditions includes the steps of inducing flow of patient blood through a blood treatment device consisting of a bioreactor inlet and outlet in fluid connection to the circulatory system of a patient. Biological agents including lipopolysaccharide (LPS) and extracellular adenosine triphosphate (ATP) contained within patient blood can be irreversibly detoxified by passage of patient blood over a bioreactor surface having attached or immobilized alkaline phosphatase enzymes and acyloxyacyl hydrolase enzyme, with the bioreactor being contained within the blood treatment device. The method uses continuous treatment of a patient's blood to convert LPS and extracellular ATP in blood into inhibitors of inflammation in vivo without adding any chemicals to the bloodstream of the patient.

The invention claimed is: 
     
       1. A blood treatment method, comprising:
 inducing flow of blood of a human or animal patient through a blood treatment device inlet and outlet; and 
 detoxifying at least one biological agent contained within the blood by passing the blood over at least one bioreactor surface having attached a phosphatase enzyme and/or an acyloxyacyl hydrolase (AOAH) enzyme, with the at least one bioreactor surface being contained within the blood treatment device and wherein the at least one biological agent is selected from the group consisting of: 
 Gram negative bacterial lipopolysaccharide (LPS); 
 Gram negative bacterial flagellin; 
 Gram positive bacterial lipoteichoic acid; 
 Gram positive bacterial flagellin; 
 one or more bacterial extracellular nucleoside triphosphates including adenosine triphosphate (ATP), nucleoside diphosphates including adenosine diphosphate (ADP), and nucleotides including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA); 
 one or more yeast extracellular nucleoside triphosphates including ATP, nucleoside diphosphates including ADP, and nucleotides including DNA and RNA; 
 one or more fungal extracellular nucleoside triphosphates including ATP, nucleoside diphosphates including ADP, and nucleotides including DNA and RNA; 
 one or more viral extracellular nucleotides including DNA and RNA; and 
 one or more host extracellular nucleoside triphosphates including ATP, nucleoside diphosphates including ADP, and nucleotides including DNA and RNA, and combinations thereof, wherein the blood treatment device includes a bioreactor associated with the bioreactor surface, and wherein the bioreactor comprises at least one of one or more surfaces of a hollow fiber bundle in a stent. 
 
     
     
       2. The blood treatment method of  claim 1 , wherein the hollow fiber bundle comprises 400 to 20,000 biocompatible hollow fibers made from polysulfone, wherein the hollow fibers have an internal diameter of 50 to 500 μm, a wall thickness of 10 to 50 μm, and a length of 3 to 50 cm, and wherein the hollow fibers are arranged in parallel in the hollow fiber bundle. 
     
     
       3. The blood treatment method of  claim 1 , wherein the phosphatase enzyme includes one or more human phosphatase enzymes including any combination of alkaline phosphatase (AP), apyrase/cluster of differentiation 39 (CD39; ecto-apyrase), and cluster of differentiation CD73 (CD73; ecto-5′-nucleotidase). 
     
     
       4. The blood treatment method of  claim 1 , wherein the acyloxyacyl hydrolase (AOAH) enzyme comprises human AOAH from human blood or tissues or synthetic human AOAH from recombinant deoxyribonucleic acid (DNA) technology. 
     
     
       5. The blood treatment method of  claim 1 , further comprising connecting the blood treatment device inlet and outlet to the patient. 
     
     
       6. The blood treatment method of  claim 1 , further comprising treating the blood continuously. 
     
     
       7. The blood treatment method of  claim 1 , further comprising circulating the blood through one or more indwelling stents and catheters by a blood pressure created by a beating heart of the patient. 
     
     
       8. The blood treatment method of  claim 1 , wherein phosphatase enzymes including, but not limited to alkaline phosphatase (AP), intestinal alkaline phosphatase (IAP),  Saccharomyces boulardii  AP (SBAP), apyrase, apyrase/cluster of differentiation 39 (CD39; ecto-apyrase), and cluster of differentiation 73 (CD73; ecto-5′-nucleotidase) irreversibly detoxify the at least one biological agent contained within the blood by dephosphorylation. 
     
     
       9. The blood treatment method of  claim 1 , wherein the AOAH enzyme detoxifies the at least one biological agent contained within the blood by deacylation. 
     
     
       10. The blood treatment method of  claim 1 , wherein one or more enzymes including alkaline phosphatase (AP),  Saccharomyces boulardii  AP (SBAP), apyrase, apyrase/cluster of differentiation 39 (CD39; ecto-apyrase), cluster of differentiation 73 (CD73; ecto-5′-nucleotidase), and AOAH enzymes are covalently attached to the bioreactor surface. 
     
     
       11. The blood treatment method of  claim 1 , wherein detoxifying the at least one biological agent contained within the blood of the patient therapeutically treats at least one of sepsis, septic shock, inflammation, bacteremia, bacterial infections, yeast infections, fungal infections, viral infections, systemic inflammatory response syndrome (SIRS), Gram negative bacterial lipopolysaccharide (LPS) in patient blood, Gram positive bacterial lipoteichoic acid in the blood, inflammatory bowel disease (IBD), inflammatory bowel syndrome (IBS), Crohn's disease, ulcerative colitis, enterocolitis, necrotizing enterocolitis (NEC), meningitis, meningococcemia, trauma or hemorrhagic shock, burns, liver disease, pancreatitis, periodontal disease, pneumonia, cystic fibrosis, asthma, alpha-1 antitrypsin (A1AT) deficiency, chronic obstructive pulmonary disease (COPD), pulmonary fibrosis, tuberculosis, coronary heart disease, congestive heart failure, infectious endocarditis, renal disease, hemolytic uremic syndrome, kidney disease, autoimmune diseases including rheumatoid arthritis, systemic lupus erythematosus, mast cell activation disorders, cancer, diabetes, infection resulting in LPS or lipoteichoic acid in patient blood, abscesses resulting in LPS or lipoteichoic acid in patient blood, protein aggregation disorders including neurodegenerative diseases, Alzheimer's disease, Parkinson's disease, Huntington's chorea, macular degeneration, amyloidosis, and amyotrophic lateral sclerosis, arthritis, atherosclerosis, aging, cancer, diabetes, obesity, emotional stress, anxiety, and patients undergoing surgery, chemotherapy and transplants. 
     
     
       12. The blood treatment method of  claim 1 , wherein the at least one bioreactor surface is provided with continuous flow of the blood that continues until the at least one biological agent being detoxified has each been reduced to predetermined levels. 
     
     
       13. The blood treatment method of  claim 1 , wherein an immobilized anti-protease plasma protein comprising α 2 -macroglobulin is used singly or in combination with one or more immobilized alkaline phosphatase (AP) and AOAH enzymes in the at least one bioreactor surface to reduce proteolytic destruction of the immobilized enzymes, to maintain activity of the enzymes in the bioreactor, and to reduce tissue destruction and inflammation caused by proteases in the bloodstream. 
     
     
       14. The blood treatment method of  claim 1 , wherein the phosphatase and AOAH enzymes are immobilized and are used to remove selected LPS and proinflammatory compounds within a biological system, including but not limited to those produced by microorganisms including bacteria, yeast, fungi, viruses and infected or damaged host tissues in humans and animals, and more specifically, to remove LPS, lipoteichoic acid, flagellin, and microbial or host extracellular ATP, ADP, DNA, and RNA in a bloodstream of the patient by passage of the blood over the bioreactor surface in a blood treatment device without adding any chemicals to the blood of the patient for continuous treatment until the LPS and proinflammatory compounds have been reduced to predetermined levels. 
     
     
       15. The blood treatment method of  claim 1 , further comprising surgically implanting an indwelling bioreactor into a patient to enable blood of the patient to flow in the following way:
 from a first vein through an inlet associated with the indwelling bioreactor and an outlet associated with the bioreactor in fluid connection to the first vein of the patient. 
 
     
     
       16. The blood treatment method of  claim 1 , wherein the dephosphorylation of extracellular ATP to adenosine monophosphate (AMP) plus phosphate by the combined action of immobilized alkaline phosphatase (AP) and the action of cluster of differentiation CD73 (CD73; ecto-5′-nucleotidase) on the endothelial surfaces of blood vessels and tissue cells of a patient results in an in vivo conversion of ATP into adenosine which inhibits inflammation without adding chemicals to the bloodstream of the patient. 
     
     
       17. The blood treatment method of  claim 1 , wherein immobilized AOAH on the at least one bioreactor surface enables in vivo conversion of proinflammatory LPS to deacylated LPS (dLPS), a competitive inhibitor of LPS for cellular attachment sites, so that dLPS inhibits inflammation caused by LPS without adding chemicals to the bloodstream of the patient.